Recombinant production, characterization and industrial application testing of a novel acidic exo/endo-chitinase from Rasamsonia emersonii.

An acid-active exo/endo-chitinase; comprising a GH18 catalytic domain and substrate insertion domain; originating from the thermophilic filamentous fungus Rasamsonia emersonii, was expressed in Pichia pastoris. In silico analysis including phylogenetic analysis, and recombinant production, purification, biochemical characterisation, and industrial application testing, was carried out. The expressed protein was identified by SDS-PAGE as a smear from 56.3 to 125.1 kDa, which sharpens into bands at 46.0 kDa, 48.4 kDa and a smear above 60 kDa when treated with PNGase F. The acid-active chitinase was primarily a chitobiosidase but displayed some endo-chitinase and acetyl-glucosamidase activity. The enzyme was optimally active at 50 °C, and markedly low pH of 2.8. As far as the authors are aware, this is the lowest pH optima reported for any fungal chitinase. The acid-active chitinase likely plays a role in chitin degradation for cell uptake in its native environment, perhaps in conjunction with a chitin deacetylase. Comparative studies with other R. emersonii chitinases indicate that they may play a synergistic role in this. The acid-active chitinase displayed some efficacy against non-treated substrates; fungal chitin and chitin from shrimp. Thus, it may be suited to industrial chitin hydrolysis reactions for extraction of glucosamine and chitobiose at low pH.

BatCount: A software program to count moving animals.

One of the biggest challenges with species conservation is collecting accurate and efficient information on population sizes, especially from species that are difficult to count. Bats worldwide are declining due to disease, habitat destruction, and climate change, and many species lack reliable population information to guide management decisions. Current approaches for estimating population sizes of bats in densely occupied colonies are time-intensive, may negatively impact the population due to disturbance, and/or have low accuracy. Research-based video tracking options are rarely used by conservation or management agencies for animal counting due to the perceived training and elevated operating costs. In this paper, we present BatCount, a free software program created in direct consultation with end-users designed to automatically count bats emerging from cave roosts (historical populations 20,000-250,000) with a streamlined and user-friendly interface. We report on the software package and provide performance metrics for different recording habitat conditions. Our analysis demonstrates that BatCount is an efficient and reliable option for counting bats in flight, with performance hundreds of times faster than manual counting, and has important implications for range- and species-wide population monitoring. Furthermore, this software can be extended to count any organisms moving across a camera including birds, mammals, fish or insects.

Pulmonary drug delivery for acute respiratory distress syndrome.

The acute respiratory distress syndrome (ARDS) is a life-threatening condition that causes respiratory failure. Despite numerous clinical trials, there are no molecularly targeted pharmacologic therapies to prevent or treat ARDS. Drug delivery during ARDS is challenging due to the heterogenous nature of lung injury and occlusion of lung units by edema fluid and inflammation. Pulmonary drug delivery during ARDS offers several potential advantages including limiting the off-target and off-organ effects and directly targeting the damaged and inflamed lung regions. In this review we summarize recent ARDS clinical trials using both systemic and pulmonary drug delivery. We then discuss the advantages of pulmonary drug delivery and potential challenges to its implementation. Finally, we discuss the use of nanoparticle drug delivery and surfactant-based drug carriers as potential strategies for delivering therapeutics to the injured lung in ARDS.

A Multi-Experiment Investigation of the Effects Stance Width on the Biomechanics of the Barbell Squat.

This two-experiment study aimed to explore habitual and manipulated stance widths on squat biomechanics. In experiment one, 70 lifters completed back squats at 70%, 1 repetition maximum (1RM), and were split into groups (NARROW < 1.06 * greater trochanter width (GTW), MID 1.06−1.18 * GTW and WIDE > 1.37 * GTW) according to their self-selected stance width. In experiment two, 20 lifters performed squats at 70%, 1RM, in three conditions (NARROW, MID and WIDE, 1.0, 1.25 and 1.5 * GTW). The three-dimensional kinematics were measured using a motion capture system, ground reaction forces (GRF) using a force platform, and the muscle forces using musculoskeletal modelling. In experiment two, the peak power was significantly greater in the NARROW condition, whereas both experiments showed the medial GRF impulse was significantly greater in the WIDE stance. Experiment two showed the NARROW condition significantly increased the quadriceps forces, whereas both experiments showed that the WIDE stance width significantly enhanced the posterior-chain muscle forces. The NARROW condition may improve the high mechanical power movement performance and promote the quadriceps muscle development. Greater stance widths may improve sprint and rapid change-of-direction performance and promote posterior-chain muscle hypertrophy. Whilst it appears that there is not an optimal stance width, these observations can be utilized by strength and conditioning practitioners seeking to maximize training adaptations.

Two-Experiment Examination of Habitual and Manipulated Foot Placement Angles on the Kinetics, Kinematics, and Muscle Forces of the Barbell Back Squat in Male Lifters.

This two-experiment study aimed to examine the effects of different habitual foot placement angles and also the effects of manipulating the foot placement angle on the kinetics, three-dimensional kinematics and muscle forces of the squat. In experiment 1, seventy lifters completed squats at 70% of their one repetition maximum using a self-preferred placement angle. They were separated based on their habitual foot angle into three groups HIGH, MEDIUM and LOW. In experiment 2, twenty lifters performed squats using the same relative mass in four different foot placement angle conditions (0°, 21°, 42° and control). Three-dimensional kinematics were measured using an eight-camera motion analysis system, ground reaction forces (GRF) using a force platform, and muscle forces using musculoskeletal modelling techniques. In experiment 1, the impulse of the medial GRF, in the descent and ascent phases, was significantly greater in the HIGH group compared to LOW, and in experiment 2 statistically greater in the 42° compared to the 21°, 0° and control conditions. Experiment 2 showed that the control condition statistically increased quadriceps muscle forces in relation to 0°, whereas the 0° condition significantly enhanced gluteus maximus, gastrocnemius and soleus forces compared to control. In experiment 1, patellofemoral joint stress was significantly greater in the HIGH group compared to LOW, and in experiment 2, patellar and patellofemoral loading were statistically greater in the control compared to the 42°, 21°, 0° and control conditions. Owing to the greater medial GRF's, increased foot placement angles may improve physical preparedness for sprint performance and rapid changes of direction. Reducing the foot angle may attenuate the biomechanical mechanisms linked to the aetiology of knee pathologies and to promote gluteus maximus, gastrocnemius and soleus muscular development. As such, though there does not appear to be an optimal foot placement angle, the observations from this study can be utilised by both strength and conditioning and sports therapy practitioners seeking to maximise training and rehabilitative adaptations.

The Effects of Sprint vs. Resisted Sled-Based Training; an 8-Week in-Season Randomized Control Intervention in Elite Rugby League Players.

The aim of the current study was to examine the efficacy of resisted sled-based training compared to traditional unresisted sprint training in terms of mediating improvements in speed, agility, and power during an eight-week period of in-season training in elite rugby league players. Participants were randomly separated into either resisted sled or traditional sprint-based training groups and they completed an eight-week in-season training block with training prescribed based on the group to which they were assigned. Measures of 5 m, 10 m, and 20 m sprint times in addition to countermovement jump height and 505-agility test time were measured at baseline, four-weeks and eight-weeks. For sprint-based outcomes, although both groups improved significantly, there were no statistical differences between the two training methods. However, at the eight-week time point there were significant improvements in 505-agility test (sprint group: baseline = 2.45 and eight-weeks = 2.42 s/sled group: baseline = 2.43 and eight-weeks = 2.37 s) and countermovement jump (sprint group: baseline = 39.18 and eight-weeks = 39.49 cm/sled group: baseline = 40.43 and eight-weeks = 43.07 cm) performance in the sled training group. Therefore, the findings from this investigation may be important to strength and conditioning coaches working in an elite rugby league in that resisted sled training may represent a more effective method of sprint training prescription.

Recombinant production and characterisation of two chitinases from Rasamsonia emersonii, and assessment of their potential industrial applicability.

Rasamsonia emersonii (previously Talaromyces emersonii) is a thermophilic filamentous fungus displaying optimum growth at 45 °C. It has a history of use in commercial food enzyme production. Its unfractionated chitinolytic secretome was partially characterised in the early 1990s; however, no individual chitinase from this source has been described in literature previously. This study describes two GH18 chitinases originating from the R. emersonii genome, expressed in the methylotrophic yeast P. pastoris. Chit1 comprises of a GH18 catalytic domain and Chit2 comprises of a GH18 catalytic domain and a chitin-binding motif at the C-terminal. The chitinases were expressed as glycoproteins. The apparent molecular weight of Chit1 was 35.8-42.1 kDa with a smearing tail associated with glyco-sidechains visible up to 72.2 kDa. This became two bands of 30.8 and 29.0 kDa upon de-glycosylation. The apparent molecular weight of Chit2 was 50.4 kDa, reducing to 48.2 kDa upon de-glycosylation. Both chitinases displayed endo-chitinase and chitobiosidase activity, temperature optima of 50-55 °C and low pH optima (pH 4.5 or lower); Chit1 displayed a pH optimum of 3.5, retaining > 60% maximum activity at pH 2.2, a pH range lower than most enzymes of fungal origin. Chit2 displayed the highest chitin-degrading ability at 3456 µmol/mg on 4-NP-triacetylchitotriose, but lost activity faster than Chit1, which displayed 403 µmol/mg on the same substrate. The predicted D values (time required to reduce the enzyme activity to 10% of its original value at 50 °C) were 19.2 and 2.3 days for Chit1 and Chit2, respectively. Thus, Chit1 can be considered one of few hyperthermostable chitinase enzymes described in literature to date. Their physicochemical properties render these chitinases likely suitable for shrimp chitin processing including one-step chitin hydrolysis and alternative sustainable protein processing and the attractive emerging application of mushroom food waste valorisation.Key points• Two GH18 chitinases originating from the industrially relevant thermophilic fungus R. emersonii were cloned and expressed in P. pastoris.• The purified recombinant chitinases showed low pH and high temperature optima and appreciable thermostability at industrially relevant temperatures.• The chitinases displayed characteristics that indicate their likely suitability to several industrial applications including sustainable alternative protein processing, food waste valorisation of commercial mushroom production and one-step shrimp chitin processing.

Effect of Velocity-Based Loading on Acceleration Kinetics and Kinematics During Sled Towing.

ABSTRACT: Bentley, I, Sinclair, JK, Atkins, SJ, Metcalfe, J, and Edmundson, CJ. Effect of velocity-based loading on acceleration kinetics and kinematics during sled towing. J Strength Cond Res 35(4): 1030-1038, 2021-Sled towing (ST) provides an external load in the form of a sled towed using a shoulder or waist harness and cord behind the athlete. Loading strategies have varied greatly between studies, and despite many investigations, there is little agreement on the optimum sled loading to develop the acceleration phase. The aim of this study was to investigate the kinetics and kinematics of velocity-based ST during the acceleration phase of sprinting. Twelve academy rugby league players performed a series of 6-m sprints in different conditions; uninhibited, 10, 15, and 20% velocity decrement (VDec). Sagittal plane kinematics and kinetic measures were examined using 1-way repeated-measures analysis of variance. Results indicated that ST affected trunk, knee, and ankle joint kinematics (p < 0.05). Peak knee flexion increased as sled loads increased (p < 0.05), which may enable athletes to lower their center of mass and increase their horizontal force application. Net horizontal and propulsive impulse measures were greater in all sled conditions (p < 0.05), which increased significantly because sled loadings were heavier. In conclusion, this study highlights the effects of differential loads to help coaches understand acute kinetics and kinematic changes to improve the planning of sprint training.

Effect of cadence selection on peak power and time of power production in elite BMX riders: A laboratory based study.

The aims of this study were to analyse the optimal cadence for peak power production and time to peak power in bicycle motocross (BMX) riders. Six male elite BMX riders volunteered for the study. Each rider completed 3 maximal sprints at a cadence of 80, 100, 120 and 140 revs · min-1 on a laboratory Schoberer Rad Messtechnik (SRM) cycle ergometer in isokinetic mode. The riders' mean values for peak power and time of power production in all 3 tests were recorded. The BMX riders produced peak power (1105 ± 139 W) at 100 revs · min-1 with lower peak power produced at 80 revs · min-1 (1060 ± 69 W, (F(2,15) = 3.162; P = .266; η2 = 0.960), 120 revs · min-1 (1077 ± 141 W, (F(2,15) = 4.348; P = .203; η2 = 0.970) and 140 revs · min-1 (1046 ± 175 W, (F(2,15) = 12.350; P = 0.077; η2 = 0.989). The shortest time to power production was attained at 120 revs · min-1 in 2.5 ± 1.07 s. Whilst a cadence of 80 revs · min-1 (3.5 ± 0.8 s, (F(2,15) = 2.667; P = .284; η2 = 0.800) 100 revs · min-1 (3.00 ± 1.13 s, (F(2,15) = 24.832; P = .039; η2 = 0.974) and 140 revs · min-1 (3.50 ± 0.88 s, (F(2,15) = 44.167; P = .006; η2 = 0.967)) all recorded a longer time to peak power production. The results indicate that the optimal cadence for producing peak power output and reducing the time to peak power output are attained at comparatively low cadences for sprint cycling events. These findings could potentially inform strength and conditioning training to maximise dynamic force production and enable coaches to select optimal gear ratios.

Impact of Harness Attachment Point on Kinetics and Kinematics During Sled Towing.

Resisted sprint training is performed in a horizontal direction and involves similar muscles, velocities, and ranges of motion (ROM) to those of normal sprinting. Generally, sleds are attached to the athletes through a lead (3 m) and harness; the most common attachment points are the shoulder or waist. At present, it is not known how the different harness point's impact on the kinematics and kinetics associated with sled towing (ST). The aim of the current investigation was to examine the kinetics and kinematics of shoulder and waist harness attachment points in relation to the acceleration phase of ST. Fourteen trained men completed normal and ST trials, loaded at 10% reduction of sprint velocity. Sagittal plane kinematics from the trunk, hip, knee, and ankle were measured, together with stance phase kinetics (third footstrike). Kinetic and kinematic parameters were compared between harness attachments using one-way repeated-measures analysis of variance. The results indicated that various kinetic differences were present between the normal and ST conditions. Significantly greater net horizontal mean force, net horizontal impulses, propulsive mean force, and propulsive impulses were measured (p < 0.05). Interestingly, the waist harness also led to greater net horizontal impulse when compared with the shoulder attachment (p < 0.001). In kinematic terms, ST conditions significantly increased peak flexion in hip, knee, and ankle joints compared with the normal trials (p < 0.05). Results highlighted that the shoulder harness had a greater impact on trunk and knee joint kinematics when compared with the waist harness (p < 0.05). In summary, waist harnesses seem to be the most suitable attachment point for the acceleration phase of sprinting. Sled towing with these attachments resulted in fewer kinematic alterations and greater net horizontal impulse when compared with the shoulder harness. Future research is necessary in order to explore the long-term adaptations of these acute changes.

The Presence of Bilateral Imbalance of the Lower Limbs in Elite Youth Soccer Players of Different Ages.

The aim of this study was to examine bilateral differences in ground reaction forces (GRF), measured during a deep squat (DS) exercise, in a population of elite youth soccer players. Bilateral muscle balance is a key component in promoting musculoskeletal health of performers, yet there is a limited evidence base investigating such imbalances in youth. Seventy-four subjects were assigned to performance groups according to chronological age (younger than 13, 14, 15, 16, 17 years). Analysis of physical maturity status revealed that very few players were classified as "early" or "late" maturers. Players completed an overhead DS exercise, as part of preseason functional movement screening. Peak GRF were assessed using a twin force plate system. Significant differences (p ≤ 0.05) were identified between right and left side peak GRF for all groups except the youngest (U13) and oldest (U17). Nondominant "sides" showed the highest levels of PGRF across all groups. The magnitude of PGRF was not significantly different both within and between groups, except for the left side in the U13 to U15 groups (p = 0.04). Results from this study show that performance asymmetry is marked in adolescence. There seems a "trigger point" during the early stage of adolescence, when bilateral imbalances become marked. These differences do seem to reduce during the later stages of adolescence. Correct attention to focussed training, designed to remediate any imbalance, is warranted in adolescent groups. This is important with respect of the key associations between bilateral asymmetry and risk of injury.

Electromyographic response of global abdominal stabilizers in response to stable- and unstable-base isometric exercise.

Core stability training traditionally uses stable-base techniques. Less is known as to the use of unstable-base techniques, such as suspension training, to activate core musculature. This study sought to assess the neuromuscular activation of global core stabilizers when using suspension training techniques, compared with more traditional forms of isometric exercise. Eighteen elite level, male youth swimmers (age, 15.5 ± 2.3 years; stature, 163.3 ± 12.7 cm; body mass, 62.2 ± 11.9 kg) participated in this study. Surface electromyography (sEMG) was used to determine the rate of muscle contraction in postural musculature, associated with core stability and torso bracing (rectus abdominus [RA], external obliques [EO], erector spinae [ES]). A maximal voluntary contraction test was used to determine peak amplitude for all muscles. Static bracing of the core was achieved using a modified "plank" position, with and without a Swiss ball, and held for 30 seconds. A mechanically similar "plank" was then held using suspension straps. Analysis of sEMG revealed that suspension produced higher peak amplitude in the RA than using a prone or Swiss ball "plank" (p = 0.04). This difference was not replicated in either the EO or ES musculature. We conclude that suspension training noticeably improves engagement of anterior core musculature when compared with both lateral and posterior muscles. Further research is required to determine how best to activate both posterior and lateral musculature when using all forms of core stability training.

The influence of different footwear on 3-D kinematics and muscle activation during the barbell back squat in males.

The barbell back squat is commonly used by athletes participating in resistance training. The barbell squat is typically performed using standard athletic shoes, or specially designed weightlifting footwear, although there are now a large number of athletes who prefer to squat barefoot or in barefoot-inspired footwear. This study aimed to determine how these footwear influence 3-D kinematics and muscle activation potentials during the barbell back squat. Fourteen experienced male participants completed squats at 70% 1 rep max in each footwear condition. 3-D kinematics from the torso, hip, knee and ankle were measured using an eight-camera motion analysis system. In addition, electromyographical (EMG) measurements were obtained from the rectus femoris, tibialis anterior, gastrocnemius, erector spinae and biceps femoris muscles. EMG parameters and joint kinematics were compared between footwear using repeated-measures analyses of variance. Participants were also asked to subjectively rate which footwear they preferred when performing their squat lifts; this was examined a chi-squared test. The kinematic analysis indicated that, in comparison to barefoot the running shoe was associated with increased squat depth, knee flexion and rectus femoris activation. The chi-squared test was significant and showed that participants preferred to squat barefoot. This study supports anecdotal evidence of athletes who prefer to train barefoot or in barefoot-inspired footwear although no biomechanical evidence was found to support this notion.

High magnetic shear gain in a liquid sodium stable Couette flow experiment: a prelude to an α-Ω dynamo.

The Ω phase of the liquid sodium α-Ω dynamo experiment at New Mexico Institute of Mining and Technology in cooperation with Los Alamos National Laboratory has demonstrated a high toroidal field B(ϕ) that is ≃8×B(r), where B(r) is the radial component of an applied poloidal magnetic field. This enhanced toroidal field is produced by the rotational shear in stable Couette flow within liquid sodium at a magnetic Reynolds number Rm≃120. Small turbulence in stable Taylor-Couette flow is caused by Ekman flow at the end walls, which causes an estimated turbulence energy fraction of (δv/v)(2)∼10(-3).